Scientists have developed a protocol for analyzing the movement and uptake of isotopically labeled azoic acid in model plants Arabidopsis thalianaproviding valuable insights into plant immune responses. This research is particularly important for understanding how plants trigger systemic resistance to pathogens. The protocol, detailed by Dr. Nicolás Cecchini, Dr. Suruchi Roychoudhry and Professor Jean Greenberg, focuses on studying a molecule that spreads within the plant, triggering tissue resistance away from the site of infection.
Professor Jean Greenberg of the University of Chicago and Dr Nicolás Cchini of the University of Córdoba pioneered the work, which was published in the journal Interstellar Protocol. The researchers devised methods for the movement of azoic acid from leaves to other parts of the plant and its uptake into the leaf disc. Professor Greenberg said: “Understanding the generation, movement, uptake and perception of mobile defense signals is key to systemic resistance programs against pathogens in flowering plants.”
The team took advantage of radioactively labeled Azelaic Acid, a version of Azelaic Acid labeled Carbon-14, to track its movement through plants. The compound was applied to a single leaf and the researchers tracked its journey to the antennae and root tissue, providing key information about how the plant mobilizes its defense signals. A noteworthy result was the movement of sulfuric acid from treated leaves to distal tissues, providing insight into how plants complement their immune responses among various organs. Additionally, controlled experiments using radiolabeled sucrose help distinguish sulfate-specific movements from general transport issues in plant-derived systems.
The researchers extended this study by analyzing azoic acid uptake into leaf discs, where they applied radiolabeled diacidic acid and observed its uptake into the tissue. They further explored the potential for root-to-shoot transport using deuterium-labeled gluconic acid, a version of aza-acid labeled with deuterium. These methods open new avenues for investigating how plants mobilize defense signals from roots to firing, providing comprehensive insights into their immune systems.
The use of isotopically labeled azoic acid has several advantages, not least its high sensitivity when detecting small numbers of molecules moving within plants. However, as Dr. Cecchini points out, there are challenges with working with radioactive materials. He explains: “We can track the movement of radiolabeled molecules, but specialized techniques such as gas chromatography-mass spectrometry (GC-MS) are needed to confirm whether the sulfuric acid remains intact or has been converted into another species during transport. form.”
The study’s findings have profound implications for plant biology, as they could improve understanding of how plants develop long-lasting resistance to pathogens. Systemic resistance is a plant immune response that requires signals that move from one part of the plant to another. When faced with subsequent infections, sulfuric acid plays a vital role in this process, resulting in a faster and more effective response.
Going forward, these protocols may be applicable to studying other plant species and defense signals. The researchers believe that their method can be used to study different small molecules involved in plant immunity, providing a powerful toolset for future research on systemic plant resistance.
Journal reference
Roychoudhry, Suruchi, Jean T. Greenberg, and Nicolás M. Cecchini. “Protocol for analysis of movement and uptake of the isotopically labeled signaling molecule sulfate in Arabidopsis.” Star Protocol (2024). doi: https://doi.org/10.1016/j.xpro.2024.102944
About the author
Jean Greenberg: Since 1997, I have been a professor in the Department of Molecular Genetics and Cell Biology at the University of Chicago. I received my degrees from Barnard College (BA, Biochemistry, Magna cum Laude) and Harvard University (PhD, Biophysics) as a Science Foundation-supported Postdoctoral Fellow in the Department of Molecular Biology at Massachusetts General Hospital/Harvard University. I have authored 76 peer-reviewed publications and hold three active patents. My work has collectively received over 14,000 Google Scholar citations, with an H-INDEX of 51 and an i10 index of 71. I was a senior editor at Plant Cell for many years. My honors include being a Pew Biomedical Scholar and a fellow of the American Society for Plant Biology. Recently, I was elected as the next president of the International Society for Molecular Plant-Microphyll Interactions. My laboratory’s contributions include: (1) Discovery and characterization of the mechanism of action of type III virulence effectors from the pathogen Pseudomonas. (2) Identification of a novel plant system defense priming signal (Azelaic, AZA) now commercially used to enhance plant health. (3) Discover the genes required for the action and movement of the system of AZA and show that these genes (Azi1 and Earli1), as well as other genes in the same family, are also required for growth and/or immune responses to promote plant growth in P. Simiae P. Simiae. and/or colonization by this strain. Signaling proteins affecting the intracellular trafficking of AZI1 and Earli1 are required to influence the multiple responses to Simiae and Aza stimulation. (4) Study metabolite and microbial peptide signaling movements to establish their systemic movements and effects. (5) Plant proteins are found to be important for basic and systemic disease resistance and cell death control. [email protected]
Nicolás M. Cecchini: I received my PhD in Chemical Sciences from the National University of Córdoba, Argentina. My PhD work elucidated the critical role of proline metabolism in controlling cell death during plant infection (Cecchini et al., 2011a, 2011b). After my PhD, I undertook postdoctoral training under Dr. Jean T. Greenberg at the University of Chicago, USA, where I investigated the mechanisms of immune memory or priming. In particular, the role of the aminotransferase ALD1 and the lipid transfer protein (LTP) AZI1 and related proteins (Cecchini et al., 2015a, 2015b, 2015b, 2019). As an independent researcher back in Argentina, I continue to explore the plastid machinery targeting AZI1 (Cecchini et al., 2020) and initiating alternative splicing of the alternative splicing driven DNA Repair/epigenetic factor MBD4L (Cecchini al Al Study of driven subnuclear localization (Cecchini et al., 2022). These studies led me to focus my current research on NLR-type immune receptors targeted to plastids and changes in chromatin and alternative splicing as mechanisms establishing the primed state (Miranda et al., 2023; Peppino et al., 2024, 2024, , 2024, in preparation).
Centro de Respeivationes enquímica Biológica De Córdoba (Ciquibic-Conicet) | Depto. química Biológica, fcq-unc.
av. Haya de la Torre S/N – Ciudad Universitaria | University of Nacordoba | CP. X5000HUA – Córdoba – República Argentina | Tel: +54 351 5353855.
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Suruchi Roychoudhry: I completed my undergraduate degree (Theoretical Biotechnology) from India, and my MSc in Molecular Biology from the University of Sussex. I then started my PhD in 2009 in the laboratory of Professor Stefan Kepinski in Plant Developmental Biology at the University of Leeds, exploring the role of the plant hormone auxin in regulating skewed growth in flowering plants role in the pattern. The results of my PhD led to the commercialization of my research and the filing of a US patent (modified plant cells), which further supported a short-lived postdoctoral project that continued with some proof-of-concept in Professor Kepinski’s laboratory Work. Next, I moved to Jean Greenberg’s laboratory at the University of Chicago (Chicago, USA), starting in 2015, to investigate the molecular mechanisms of systemic plant immunity. I then moved back to Leeds in 2016 and after taking a break from my parents’ time as a parent and returning to work part-time, I am currently a senior researcher at the Center for Plant Science at the University of Leeds, investigating angle-dependent gravity responses in Arabidopsis roots. Mechanism, while applying for (and being rejected) (and being rejected) from! ) Independent Scholarship Position. Twitter: @Suruchiroy1 Email: [email protected]
